摘要
采用改进的碱裂解法提取Gluconacetobacter hansenii ATCC23769自发不产膜突变体的内源隐蔽质粒。用不同的限制性内切酶对混合质粒直接进行酶切,酶切后的片段混合物与pUC18载体连接构建重组载体。重组载体回转入G.hansenii ATCC23769获得隐蔽质粒上具有复制能力的片段,序列结果分析表明:该片段上没有某些其他质粒所具有的Rep蛋白。利用该片段,构建了能同时在大肠杆菌和葡糖酸醋杆菌中复制的质粒载体,体内的抗生素抗性实验证明该载体具有良好的稳定性。
The endogenous cryptic plasmids of Gluconacetobacter hansenii ATCC23769 spontaneous cellulose-negative were extracted by modified alkaline-lysis method.The plasmids were directly digested by certain restriction endonucleases,and the digested fragments were ligated to pUC18 vector to construct the recombinant vectors.The recombinant vectors were re-transformed into G.hansenii ATCC23769 to obtain the replication fragment of the cryptic plasmids.The sequence analysis showed that this fragment did not contain the Rep protein.The vector replicated in both E.coli and Gluconacetobacter was constructed,and its good stability was confirmed in antibiotic-resistant experiments.
出处
《生物加工过程》
CAS
CSCD
2011年第3期47-51,共5页
Chinese Journal of Bioprocess Engineering
基金
福建省科技项目(2005Q007)
关键词
葡糖酸醋杆菌
混合质粒
复制
Gluconacetobacter
plasmid mixture
replication
